LOCUS       sequence-405417-        2686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Receiver cloning vector for even-3 level constructs. Contains lacZ 
            for blue/white screening..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2686)
  AUTHORS   Pollak B, Matute T, Nunez I, Cerda A, Lopez C, Vargas V, Kan A, 
            Bielinski V, von Dassow P, Dupont CL, Federici F
  TITLE     Universal loop assembly: open, efficient and cross-kingdom DNA 
            fabrication.
  JOURNAL   Synth Biol (Oxf). 2020;5(1):ysaa001. doi: 10.1093/synbio/ysaa001. 
            Epub 2020 Feb 5.
  PUBMED    32161816
REFERENCE   2  (bases 1 to 2686)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Synth Biol 
            (Oxf)."; date: "2020"; volume: "5(1):ysaa001. doi"; pages: " 
            10.1093/synbio/ysaa001. Epub 2020 Feb 5"
COMMENT     Sequence Label: pCA_e-3
FEATURES             Location/Qualifiers
     source          1..2686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    38..54
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     43..65
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     62..78
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     62..78
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             74..397
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    91..147
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(148..165)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(148..164)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(157..179)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(373..392)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(655..1243)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(735..754)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1334..2125)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     protein_bind    2650..2671
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2686,1..5,6..23,24..30)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
       61 acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga
      121 ggatccccgg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga
      181 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
      241 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
      301 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
      361 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
      421 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
      481 tgtgacacta gtgagcacgc tcttcaggta gctttgagac caccccagga tacatagatt
      541 accacaactc cgagcccttc cacccacaga atcaggggat aacgcaggaa agaacatgtg
      601 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      661 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      721 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
      781 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
      841 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
      901 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
      961 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1021 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1081 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1141 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1201 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1261 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgca
     1321 ttctaggtga ttattatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa
     1381 attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct
     1441 gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg
     1501 cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg
     1561 taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg
     1621 tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg
     1681 ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa
     1741 tgtcgatcgt ggctggctcg aagataccag caagaatgtc attgcgctgc cattctccaa
     1801 attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg
     1861 tgacttctac agcgcggaga atctcactct ctccagggga agccgaagtt tccaaaaggt
     1921 cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat
     1981 caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca
     2041 gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata
     2101 cttcggcgat caccgcttcc ctcatgcgaa acgatcctca tcctgtctct tgatcagata
     2161 ttgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg
     2221 gcttcccaac cttaccagag ggcgccccag ctggcaattc cggttcgctt gctaagcttg
     2281 catgcctgca agtcgactct agaggagcgg tgatcacagg cagcaacgct ctgtcatcgt
     2341 tacaatcaac atgctaccct ccgcgagatc atccgtgttt caaacccggc agcttagttg
     2401 ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca acggctctcc
     2461 cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt gtgccgagct
     2521 cattactcgc atccattctc aggctgtctc gtctcgtctc cgaggtctca aatgaatgtg
     2581 aagagcttga cagcgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     2641 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggct
//
